AbVideo™ - Antibody Conjugation - FITC

The conjugation of polyclonal and monoclonal antibodies with fluorescein isothiocyanate (FITC) is used in immunohistochemistry and immunofluorescence studies. FITC isomer I is a widely used fluorescent labeling reagents due to the fluorophores high quantum efficiency and conjugate stability.

alamarBlueR works as a cell viability and proliferation indicator through the conversion of resazurin to resorufin. Resazurin, a non-fluorescent indicator dye, is converted to highly red fluorescent resorufin via reduction reactions of metabolically active cells. The amount of fluorescence produced is proportional to the number of living cells.

Alizarin Red is used to identify calcium deposits in tissue sections. Calcium forms an Alizarin Red S-calcium complex in a chelation process. This video describes the procedure of Alizarin Red S Staining for osteogenesis.

Antibodies labeled with biotin provide the user with a tool for increasing the sensitivity of an assay by its ability to amplify a given reaction. A biotin/streptavidin (avidin) system offers low background, enhanced sensitivity and the ability to detect minute amounts of antigens.

The in situ proximity ligation assay is a powerful technology capable of detecting single protein events such as protein protein interactions (e.g. protein dimerization) and modifications (e.g. protein phosphorylation) in tissue and cell samples prepared for microscopy. Each detected signal is visualized as an individual fluorescent dot, these signals can be quantified (counted) and assigned to a specific subcellular location based on microscopy images.

Cyanogen bromide (CNBr) is the most common method for preparing affinity chromatography to purify antibody because of its simplicity and mild pH conditions. CNBr reacts with the hydroxyl groups on agarose to form cyanate esters and imidocarbonates. These groups are reacted with primary amines in order to couple the protein onto the agarose matrix.

Protein A Sepharose is prepared by covalently coupling Protein A to 6% cross-linked sepharose beads. The coupling technique is optimized to give a high binding capacity for IgG. This protocol is a simple, reliable method for purifying total IgG from crude protein mixtures such as serum or ascites fluid.

Polyethylene Glycol (PEG) is a long chain polymer that has been approved by the Food and Drug Administration for human intravenous, oral and dermal applications. Covalent attachment of PEG (PEGylation) to proteins can reduce their immunogenicity, minimize proteolytic cleavage and increase their serum half-life. PEG has also been attached to small molecules and liposomes for more selective delivery. PEG-modification of superparamagnetic iron oxide and quantum dots can improve their biocompatibility and reduce non-specific uptake. This video shows the procedure of sandwich ELISA assay for anti-PEG antibody pair.

The ApoTox-Glo™ Triplex Assay combines three Promega assay chemistries to assess viability, cytotoxicity and apoptosis within a single assay well. The first part of the assay simultaneously measures two protease activities: GF-AFC is used to measure cell viability, and bis-AAF-R110 to cytotoxicity. The second part of the assay uses the Caspase-GloR to measure caspase activity.

Besides the temperature logger, self-contained biological indicators that contain G. stearothermophilus spores can also be used for autoclave quality control test. Learn how to use the biological indicator in this AbVideo. There are also physical and chemical indicators that can be used to ensure decontamination effectiveness of autoclaves.

Stainless steel loggers are commonly used for monitoring temperature of autoclave. Check out how to use it to maintain the maximum performance of your autoclave machine. There are physical, chemical, and biological indicators that can be used to ensure decontamination effectiveness of autoclaves.

Biological Safety Cabinets (BSC) is an enclosed, ventilated laboratory workspace which provides the most effective primary containment for working with infectious agents. The BSC is tested for bioburden to ensure the adequate protection.

The BioSpectrum® Imaging System is designed to automate the research with one-touch, pre-set or user-defined controls for accurate, repeatable imaging and analysis for chemiluminescent, bioluminescent, fluorescent, colorimetric imaging. The BioSpectrum® is faster than film for western blot imaging.

A blood smear gives information about the number and shape of blood cells. To perform a blood smear, two very clean slides are required. Drop a small drop of blood on one of the slides. Pull blood forward across slide.

Calcium phosphate transfection method is a simple, time efficient and relatively inexpensive way to introduce DNA into cells in stable cell lines such as HEK293T, CHO and BHK cells. This method works best in cell lines that are highly transformed and adherent and this technique requires few manipulative steps and maintains high levels of reproducibility from experiment to experiment.

jetPEI™, a polyethylenimine (PEI) derivative, is a cationic polymer transfection reagent that binds to DNA by electrostatic forces. The cationic polymer/DNA complex enters the cell by endocytosis, where the DNA is then released into the cytoplasm.

The cells must be fixed and permeabilized to ensure free access of the antibody to its antigen. Fixation methods fall generally into two classes: organic solvents and cross-linking reagents. This video uses formaldehyde as a cross-linking reagent to fix cells.

CellTiter 96R AQueous One Solution Cell Proliferation Assay is a colorimetric method for determining the number of viable cells in proliferation or cytotoxicity assays. The reagent contains a tetrazolium compound (MTS). The MTS is bioreduced by cells into a colored formazan product which can be measured by the absorbance at 490nm.

The Cell Proliferation Reagent WST-1 is a clear, slightly red, ready-to-use solution, containing WST-1, a tetrazolium salt. By cellular mitochondrial dehydrogenases, WST-1 is bioreduced into formazan which can be measured at an absorbance of 440 nm.

Cell Synchronization is a process by which cells at different stages of the cell cycle in a culture are brought to the same phase. Nocodazole is one of anticancer drugs that synchronize cells at M-phase. It can interfere with the structure and function of microtubules in interphase and mitotic cells.

Chimera RNAi is a process by which small interfering RNA/DNA chimera triggers the destruction of mRNA. The discovery work, design, and application of chimera RNAi have been pioneered by Professor Kaoru Saigo and Dr. Kumiko Ui-Tei at the University of Tokyo.

Clonogenic assay allows one to test the capability of adherent cells to survive and replicate following insult with chemicals or radiation. Count the crystal violet stained colonies which incubated for 9 days with appropriate chemical or radiation dose and calculate the survival rate.

The Fyrite gas analyzer is the most widely used instrument for measuring carbon dioxide (CO2) levels in incubators. It uses the Orsat method of volumetric analysis involving chemical (potassium hydroxide) absorption of CO2 gas.

Competent cells are those that possess more easily altered cell walls that DNA can be passed through easily. Because DNA is hydrophilic, it will not normally pass through membrane. In order to make these cells readily incorporate foreign DNA, they must first be made "competent" to take up foreign DNA.

In competitive ELISA, unlabeled antibody is incubated in the presence of its antigen. Then these bound antibody/antigen complexes are then added to an antigen coated well. After washing, unbound antibodies are removed. The more analytes in the sample, the less antibodies will be able to bind to antigens in the well. The signal is then detected using labeled secondary antibodies and the decrease in signal is compared to a control. The major advantage of a competitive ELISA is the ability to use crude or impure samples and still selectively bind any antigen that may be present.

Diethylpyrocarbonate (DEPC) is used in the laboratory to inactivate the RNase enzymes from water and other laboratory utensils. It inactivates the RNases by the covalent modifications of the histidine residues. DEPC treated (and therefore RNase-free) water is used in handling of RNA to reduce the risk of RNA being degraded by RNases.

Human umbilical vein endothelial cells (HUVEC) are commonly used as a laboratory model system for the physiological and pharmacological investigations. This video describes how to derive HUVEC from the endothelium of veins of the umbilical cord.

Dialysis (ultrafiltration) is a method to concentrate protein or other macromolecules through a membrane with defined pores. The membranes will have a molecular weight cut-off (MWCO). This is the limit of size (or range) of a protein that can fit through the membrane.

Dot blot is a technique can be used as a semiqualitative method for rapid screening of a large number of samples. It is for detecting, analyzing, and identifying proteins, similar to the western blot technique but differing in that protein samples are not separated electrophoretically but are spotted through circular templates directly onto the membrane.

Electroporation applies a large electric pulse temporarily disturbs the phospholipid bilayer, allowing molecules to pass into the cell. It is usually used in molecular biology as a way of introducing some substance into a cell, such as molecular probes, drugs or pieces of coding DNA.

Emulsification is a process to mix adjuvants and immunogen. Adjuvants are mixed and injected with an antigen to prevent catabolism and help increase the immune response by localizing the antigen for an extended time and attracting the appropriate cells (T cells, B cells and APC) to interact with it.

Ethidium Bromide (EtBr) is commonly used as a non-radioactive marker for identifying and visualizing nucleic acid bands in electrophoresis. It is a potent mutagen, so its hazardous properties require special safe handling and disposal procedures.

A fluorescent microscope is an optical microscope used to study properties of substances using the phenomena of fluorescence and phosphorescence. Specimen can be labeled specifically with a fluorescent molecule and be illuminated with specific wavelength which is absorbed by the fluorophores.

GST fusion proteins allow convenient affinity purification of many proteins of interest but the GST-tag may become inconvenient in various downstream applications. We use PreScission protease to digest the GST fusion proteins and the protein without GST-tag is obtained.

Protein purification is a series of processes intended to isolate a single type of protein from a complex mixture. We use GST-fusion protein to purify and detect proteins of interest. The GST-fusion protein can be purified from cells via its high affinity for glutathione.

As a mature diagnostic tool, regular blood test is widely adopted to evaluate anemia, leukemia, reaction to inflammation and infections, etc. With automated hematology analyzer, the test result of CBC with DIFF (Complete Blood Count with Differential) can easily be obtained. The test collects information including the number and types of different types of blood cells, the variation in the size of red blood cells, hematocrit, hemoglobin value, platelet count, mean corpuscular hemoglobin, and the average size of red blood cells. Here we demonstrate a simple operation of hematology test for following clinical diagnosis.

Proteins with histidine tag can be purified and detected easily because the string of histidine residues binds to several types of immobilized metal ions, including nickel, cobalt and copper, under specific buffer conditions. This video provides generic protocol to purify 6xHis-tagged protein by Nickel nitrilotriacetic (Ni-NTA) sepharose.

Immunofluorescence is a technique to visualize a specific protein or antigen in cells or tissue sections by binding a specific antibody chemically conjugated with a fluorescent dye. We use the indirect immunofluorescence staining to perform cells fixed on slides and examine under a fluorescence microscope.

Immunoprecipitation (IP) is the technique of precipitating an protein antigen out of solution using an antibody specific to that antigen. This antibody is immobilized on a solid-phase substrate such as protein A/G agarose beads. The beads are then added to the protein mixture and the proteins that are targeted by the antibodies are captured onto the beads.

The indirect ELISA is used primarily to determine the strength and/or amount of antibody response in a sample. In the assay, the antigen of interested is immobilized by direct adsorption to the assay plate. Detection of the antigen can then be performed by using a matched set of primary antibody and conjugated secondary antibodies.

The CELLine bioreactor is a disposable, two-compartment cultivation device suitable for many cell culture applications. There are two sizes of the CELLine, CL350 and CL 1000. We use CL350 to produce monoclonal antibodies on a laboratory scale.

Cell invasion assay is designed to accelerate the screening process for compounds that influence cell migration through extracellular matrices. Matrigel (BD), extracted from the Engelbreth-Holm-Swarm (EHS) mouse sarcoma, resembles the extracellular matrices is used by cell biologists as a substrate in cell invasion assay.

Isotopic labeling is a technique for labeling a substance with a stable or radioactive isotope. By measuring the radioactivity or the abundance of the isotope, we could made observations of the course through a biologic process. This video shows the procedure for using radioisotope phosphorus-32 to label DNA.

Firefly luciferase is widely used as a reporter to study gene expression and other cellular events. The Luciferase Assay is an extremely sensitive and rapid reagent for quantitation of firefly luciferase. Through the assay, light is produced by converting the chemical energy of luciferin oxidation and can be measured by a luminometer.

Biological materials often are dried to stabilize them for storage or distribution. Lyophilization also called freeze-drying, is one method of drying biological materials that minimizes damage to its internal structure.

MaxBead Protein A/G is designed as a rapid and simple tool for immunoprecipitation, purification/depletion assays, and other magnetic separation applications. Antibody can easily bind to the magnetic beads due to its high affinity with
protein A/G.

The migration assay (also known as the Boyden Chamber Assay) is a commonly used test to study the migratory response of endothelial cells. During this assay, cells are placed on the upper layer of a chamber and a solution containing the test agent is placed below the chamber. Following an incubation period, the cells migrated through the membrane are stained and counted.

This procedure to isolate mononuclear cells from whole blood is based on density differences between mononuclear cells and other elements found in the blood sample. Differential migration during centrifugation results in the formation of layers containing different cell types. Mononuclear cells are at the interface between the plasma and the ficoll layer and are recovered by washing steps.

Genotyping is the process of determining the genes (genotype) of an individual by examining its DNA with the use of biological assays (ex: PCR, DNA sequencing). Three steps to type the mouse genotype : (i) isolate the genomic DNA from mouse tails; (ii) PCR to amplify with specific primers; (iii) electrophoresis to determine the genotype.

A demonstration on the procedure of using MTT assay to assess the viability and the proliferation of regular cells with absorbance detection at 595nm is shown. This colorimetric assay measures the reduction of yellow 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) by mitochondrial succinate dehydrogenase.

Bioresource Collection and Research (BCRC) provided this video to introduce the protocol of MTT assay. MTT assay allows assessing the viability and the proliferation of cells. This is a colorimetric assay that measures the reduction of yellow 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) by mitochondrial succinate dehydrogenase.

Mycoplasma is highly infectious and cross-contamination commonly occurs when new cells are introduced into laboratories. We use PCR-based method for detection of Mycoplasma weekly. It is a highly sensitive, specific and rapid method to detect mycoplasma contamination in cell cultures.

Mycoplasma is a common and serious contamination of cell cultures. Mycoplasma contamination can cause the alteration of the phenotypic cell characteristics. PCR method allows for fast and reliable identification of mycoplasma contamination in cell cultures.

This AbVideo demonstrates the operation of automated fiber cleavage to produce flat and angled cleaves on fibers ranging from 80 microns to 1.25 mm in diameter for following biological applications. Optical fibers are flexible, transparent fibers made of high quality extruded glass (silica) or plastic. It is frequently adopted as a waveguide, or "light pipe", to transmit the optical signals. Taking advantages of this kind of property, optical fibers have been used in developing biological sensors in concert with traditional bio-probes such as nucleic acids.

This AbVideo demonstrates the operation of automated interferometric inspection system specifically designed for checking the surface quality and flatness of cleaved, polished or lensed fibers. Optical fibers are flexible, transparent fibers made of high quality extruded glass (silica) or plastic. It is frequently adopted as a waveguide, or "light pipe", to transmit the optical signals. Taking advantages of this kind of property, optical fibers have been used in developing biological sensors in concert with traditional bio-probes such as nucleic acids.

Propidium iodide (or PI) is an intercalating agent and a fluorescent molecule that can be used to stain DNA. PI is used as a DNA stain for both flow cytometry to evaluate cell viability or DNA content in cell cycle analysis and microscopy to visualize the nucleus and other DNA containing organelles. It can be used to differentiate necrotic, apoptotic and normal cells.

FUJIFILM FLA-2000 is a multiple imaging application suitable for fluorescence and radioisotopic measurement. Laser scanning with the FLA-2000, instead of using conventional photography, permits analysis and digital storage. With FUJIFILM Imaging Plate (IP), a technology which is 100 times more sensitive to radiation than X-ray film, exposure times are reduced to 1/10 - 1/20 of film and amount of radioisotope is reduced.

The ability of cell migration is commonly used for the evaluation of cell invasion or wound repair. Using boyden chamber is convenient for the assessment of cells responses toward genetic manipulation or chemo attraction treatment.

RNA interference is a gene-silencing technique used in studying the absence of normal gene action. To better understand its function and role in disease, you can transfect siRNA into cell line to turn gene expression off or knock it down.

Sandwich ELISA is performed to determine the amount and serological class of antibodies made by an immunized animal or present in the serum of patients. The anti-immunoglobulin antibodies used have high specificity and sensitivity.

A serial dilution is a series of simple dilutions which amplifies the dilution factor quickly beginning with a small initial quantity of material. The dilution factor at each step is constant, resulting in a geometric progression of the concentration in a logarithmic fashion.

Tissue embedding is a tissue preparation technique. The tissue samples are embedded in embedding material (such as agar, gelatine, or wax) and ready to be sliced. This video describes how tissue embedding.

The tissue grinder provides rapid and efficient disruption of up to 12 biological samples, including animal and human tissues, plant tissues, bacteria, and yeast. Disruption and homogenization are achieved through the beating and grinding effect of beads on the sample material as they are shaken together in 2 ml sample tubes.

Tissue Microarray is a powerful new technology for high throughput analysis of protein expression in a large number of tissue samples. Hundreds of tissue cores are arranged on a single slide, and then analyzed by immunohistochemistry staining. We offer a large collection of immmunohistochmistry (IHC) validated antibodies for the tissue microarray platform.

This AbVideo demonstrates how to prepare as ready-to-use denatured lysate in 1x sample buffer. Transfected lysate is useful as a positive control in western blot and the full-length protein construct is expressed in HEK293T cells.

This AbVideo shows the steps involved on preparation of ready-to-use, native lysate in modified RIPA buffer. Transfected lysate is useful as a positive control in western blot and the full-length protein construct is expressed in HEK293T cells.

Urinalysis is one of the most common methods in medical diagnosis. It includes an array of tests performed with urine, such as the concentration of ions, trace metals, protein and other molecules. In principle, the test result of test strips can be read as color changes. With automated urine analyzer, artifacts of data judgment can be further avoided. Here is a brief demonstration of regular urine test using urine test strip in concert with automated analyzer.

Wright and Giemsa stains are used to stain peripheral blood and bone marrow smears. The components are oxidized eosin Y, methylene blue, and azure B. They stain the cytoplasm of cells an orange to pink color and nucleus a blue to purple. Wright and Giemsa stains are used to study blood cell morphology.